Patient Details

Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

PRE- IMPLANTATION GENETIC SCREENING TEST REPORT

 SCREENING RESULTS
Number of Embryo 12
Date of Biopsy (Sample type) 00/00/2023 (Trophectoderm Biopsy)

Method: Next Generation Sequencing using Ion ReproseqTM- PGS Kit Next Generation
Sequencing (NGS) was employed for aneuploidy screening with the help of Ion Reproseq TM
PGS kit on S5 GeneStudio platform. Complete workflow is four major segments viz. Library
preparations, Template preparation, Sequencing and Analysis. On receipt of the biopsy, samples
are individually barcode and labelled for further identification. Biopsy are subjected to Whole
Genome Amplification (WGA) by SingleSeqTM library preparation, followed by purification
and quantification of library using Qubit 4TM. Data were analyzed using Ion reporter and report
was generated following standard guideline.

Results: NGS analysis of the Embryo revealed the following results.

 Patient Details
Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

Sr Sample Id
Result Interpretation Mtdna/ Transfer
No. (Label on Vial)
Auto Ratio Priority

1 175/0123 Embryo Chaotic Abnormal/ 0.0003 -

DD01 Abnormality Aneuploid

2 175/0123 Embryo Normal/ Normal/ 0.0007 2

DD02 Euploid Euploid

3 175/0123 Embryo Trisomy 15 Abnormal/ 0.0008 -

DD03 (15q11.2q26.3x3.0) Aneuploid

4 175/0123 Embryo Monosomy 21 Abnormal/ 0.0004 -

DD04 (21q11.2q22.3x1.0) Aneuploid

5 175/0123 Embryo Normal/ Normal/ 0.0003 1

DD05 Euploid Euploid

6 175/0123 Embryo Normal/ Normal/ 0.0009 3

DD06 Euploid Euploid

7 175/0123 Embryo Monosomy 16 Abnormal/ 0.0006 -

DD07 (16p13.3q24.3x1.0) Aneuploid
 Patient Details
Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

8 175/0123 Embryo Mosaic Trisomy Abnormal/ 0.0008 -

DD08 11 (60%) Aneuploid
(11p15.5q25x3.0)

9 175/0123 Embryo Trisomy 21 Abnormal/ 0.0003 -

DD09 (21q11.2q22.3x3.0) Aneuploid

10 175/0123 Embryo Normal/ Normal/ 0.0011 4

DD10 Euploid Euploid

11 175/0123 Embryo Normal/ Normal/ 0.0013 5

DD11 Euploid Euploid

12 175/0123 Embryo Monosomy 4 (35%) Abnormal/ 0.002 -

DD12 & Monosomy 6 Aneuploid
(40%)
(4p16.3q35.2x1.65,
6p25.3q27x1.6)
 Patient Details
Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

Whole Genome View:

Analysis 1, Proband: Sample_DD02

Analysis 2, Proband: Sample_DD05

Analysis 3, Proband: Sample_DD06

 Patient Details
Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

Analysis 4, Proband: Sample_DD10

Analysis 5, Proband: Sample_DD11

 Patient Details
Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

Interpretation of the test:

The ion Reporter software create graph representing the chromosome copy number variations (CNV) of the
sample analyzed when compared to the reference bioinformatics baseline from the database of multiple
normal samples.

Euploid/ Normal:

An embryo is considered Euploid/Normal when the graph shows no deviation from the reference baseline
for any of the 24 chromosomes.

Aneuploid/ Abnormal:

An embryo is call abnormal or aneuploidy when there is a full aneuploidy (Gain or loss of any of the whole
chromosome 1:22 autosomes and X, Y) or partial aneuploidy (Gain or loss of segment of chromosome arm
P;q), is detected as deviation from the reference bioinformatics baseline with points shifted upwards Gain (+)
and downwards for Loss (-).

Complex Aneuploid:

An embryo is called Complex aneuploid when 2-5 aneuploidy is detected in the given sample.

Chaotic Aneuploid:

An embryo is considered chaotic when more than 5 aneuploidies are detected in the sample.
 Patient Details
Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

No DNA/ QC Fail:

Usually when no DNA is detected the cell didn’t have an intact nucleus. other factors for no DNA are poor
embryo quality and degraded DNA. In some cases, the cells are lost in washing procedure or may not be
successfully loaded in the buffer and stick to the wall of the tube.

No CALL:

When a reliable result cannot be achieved with high confidence No call will be reported for the embryo.
PGT A also analyze sex chromosome along with other 22 autosomes, the gender status of the analyzed
embryos will never be revealed in the report or in any way to patient or clinician. Sex selection/Sex
determination is prohibited and punishable in India under PC & PNDT act no. 57 1994.

TEST LIMITATION:

The test cannot detect chromosome loss / gain below 6 MB. Structural rearrangement in which there is a
balance amount of genetic material is involved, low level of mosaicism, uniparental disomy (both copies of
the chromosome inherited from a single parent), defect which affect complete set of chromosomes i.e
triploidy and single gene mutation such as sickle cell, cystic fibrosis disease.
The ratio of mtDNA and autosomes which is considered as reference to select the priority transfer of
embryos has not been clinically proven yet.

NOTE:

Validation study performed at sterling Accuris shows there is approximately 1-2% risk of misdiagnosis either
as false positive or false negative result with preimplantation genetic testing for aneuploidies. The transfer
of normal/Euploid embryo is believe to reduce the risk of pregnancy with birth defect, however there is a 3-
4 % risk in the general population for birth defect caused by genetic and /or non-genetic factors not detected
by the test. PGT A cannot be considered the replacement of prenatal diagnosis, even after doing PGT A we
recommend to do routine scan and test and follow-ups.
 Patient Details
Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

Disclaimer:

 The report represents only the Copy Number Variations / Microdeletion based on the
overall median tiling spacing with 1 MB Refseqgenes.
 The mutations have not been validated by PCR, Q-PCR or Sanger sequencing detection
method.
 Sex chromosome cannot be revealed as per PC and PNDT act.

References:

1. Fiorentino, Francesco, Anil Biricik, Sara Bono, Letizia Spizzichino, Ettore Cotroneo,
Giuliano Cottone, Felix Kokocinski, and Claude-Edouard Michel. "Development and
validation of a next-generation sequencing–based protocol for 24-chromosome aneuploidy
screening of embryos." Fertility and sterility 101, no. 5 (2014): 1375-1382.
2. Knapp, Michael, Mathias Stiller, and Matthias Meyer. "Generating barcoded libraries for
multiplex high-throughput sequencing." Ancient DNA: Methods and protocols(2012):155-
170.
3. Quianlan, Aaron R., and Ira M. Hail. “BEDTools: a flexible suite of utilities for comparing
genomic features” Bioinformatics 26, no 6 (2010): 841-842.
4. Diez-Juan A, Rubio C, Marin C, Martinez S, Al-Asmar N, Riboldi M, et al. Mito-chondrial
DNA content as a viability score in human euploid embryos: less is better. Fertil Steril
2015;104:534–41.e1.
5. Ravichandran K, McCaffrey C, Grifo J, Morales A, Perloe M, Munne S, et al.
Mitochondrial DNA quantification as a tool for embryo viability assessment: retrospective
analysis of data from single euploid blastocyst transfers. Hum Reprod 2017;32:1282–92.
 Patient Details
Patient ID 0223175000123 Clinician Name DR. XYZ

Patient Name ABC

Date of Collection 00/00/2023 Report Generated 00/00/2023
Patient Age 00 YEARS Test Requested PGS

6. Ferrick L, Lee YSL, Gardner DK. Metabolic activity of human blastocysts correlates with
their morphokinetics, morphological grade, KIDScore and artificial intelligence ranking.
Hum Reprod 2020;35: 2004–16.
7. Van Blerkom J. Mitochondrial function in the human oocyte and embryo and their role in
developmental competence. Mitochondrion 2011;11:797–813.
8. Cecchino GN, Garcia-Velasco JA. Mitochondrial DNA copy number as a predictor of
embryo viability. Fertil Steril 2019;111:205–11.

Dr. Gaurav Shastri

(Consulting Geneticist)

*** END OF THE REPORT***